Cancer has been one of main causes of human deaths for many years. Early detection of cancer is essential to provide definitive treatment. Among many cancer treatment methods, nanoparticle-mediated photothermal therapy is considered as one of the promising cancer treatment methods because of its non-invasiveness and cancer-specific therapy. Ultrasound and photoacoustic imaging can be utilized for both cancer detection and photothermal therapy guidance. Ultrasound elasticity imaging can detect cancer using tissue elastic properties. Once cancer is diagnosed, spectroscopic photoacoustic imaging can be used to monitor nanoparticle delivery before photothermal therapy. When nanoparticles are well accumulated at the tumor, ultrasound and photoacoustic-based thermal imaging can be utilized for estimating temperature distribution during photothermal therapy to guide therapeutic procedure. In this dissertation, ultrasound beamforming, elasticity imaging, and spectroscopic photoacoustic imaging methods were developed to improve cancer detection and therapy guidance. Firstly, a display pixel based synthetic aperture focusing method was developed to fundamentally improve ultrasound image qualities. Secondly, an autocorrelation based sub-pixel displacement estimation method was developed to enhance signal-to-noise ratio of elasticity images. The developed elasticity imaging method was utilized to clinically evaluate the feasibility of using ultrasound elasticity imaging for prostate cancer detection. Lastly, a minimum mean square error based spectral separation method was developed to robustly utilize spectroscopic photoacoustic imaging. The developed spectroscopic photoacoustic imaging method was utilized to demonstrate ultrasound and photoacoustic image-guided photothermal cancer therapy using in-vivo tumor-bearing mouse models. The results of these studies suggest that ultrasound and photoacoustic imaging can assist both cancer detection and therapy guidance. / text

Cancer currently represents one of the greatest burdens on human health in the world, claiming in excess of 7 million lives a year worldwide. Advances in both our understanding of the disease as well as our ability to diagnose it before it has had a chance to metastasize will lead to a reduction in its burden on society. To these ends, optical imaging techniques are particularly attractive. The ability to resolve cellular details noninvasively is paramount to improved cancer detection and to research on diseased tissue and cells. Lanthanoid nanoparticles, a group of photoluminescent contrast agents developed within the last two to three decades, have numerous unique optical properties that enable their use in improved and novel optical techniques. They possess large Stokes and anti-Stokes shifts, sharp electronic transitions, long luminescence lifetimes, and exceptional photostability. For these reasons, they are a good choice for biomedical applications that benefit from low background fluorescence or long illumination times. The major goal of the research presented in this dissertation was to synthesize functional lanthanoid nanoparticles for optical imaging modalities, and to explore their potential uses in a variety of biomedical applications. To this end, the research can be broken up into three specific aims. The first aim was to successfully and reproducibly synthesize downconverting and upconverting lanthanoid nanoparticles, and to functionalize these nanoparticles for use in optical techniques that would aid in the research and diagnosis of cancer. The second aim was to conduct a thorough investigation of the optical properties of these nanoparticles, and the third aim was to explore the utility of these nanoparticles in a variety of biomedical applications. First, both downconverting and upconverting lanthanoid nanoparticles were synthesized using several different methods, resulting in nanoparticles of varying size and surface functionality. Novel methods were employed to improve the utility of these nanoparticles for specific applications, including the incorporation of a mixed surface ligand population in downconverting lanthanoid nanoparticles and the use of a biomimetic surface coating to render upconverting nanoparticles water dispersible. These coated particles were further functionalized by the addition of folic acid and an antibody for epidermal growth factor receptor, both of which bind to cell surface receptors overexpressed in a number of cancers. Second, the spectral properties of lanthanoid nanoparticles were explored in detail, with special attention paid to many of the unique optical properties of upconverting lanthanoid nanoparticles. This included the discovery of one such unique property, the excitation frequency dependent emission of NaYF₄ nanocrystals codoped with Yb³⁺ and Er³⁺. Third, lanthanoid nanoparticles were used as contrast agents in a number of biomedical applications, including the development of a homogenous assay based on diffusion enhanced luminescence resonance energy transfer, a wide-field luminescence lifetime microscope, and a super resolution microscope based on the aforementioned excitation frequency dependent emission of NaYF₄:Yb³⁺,Er³⁺ nanoparticles. Specific binding of functionalized upconverting lanthanoid nanoparticles was investigated with laser scanning multiphoton microscopy, and an image processing technique was developed to overcome the challenge of working with long lived luminescent contrast agents using this imaging modality.

In this thesis we describe two separate experiments involving the use of gold nanoparticles. The first experiment looks at the use of gold nanoparticles as a localized heat source and the potential application as a cancer treatment. The second experiment, which is the real focus of this thesis, applies gold nanoparticles in the study of the free surface dynamics of glassy thin films. Gold nanoparticles have the ability to strongly absorb the energy in an incident laser beam and convert that energy into heat. Photothermal therapy is a proposed cancer treatment which exploits this ability to irreparably damage cancerous tissues surrounding gold nanoparticles. In the first chapter of this thesis we explain an experiment designed to probe the local temperatures achieved in such a process. Gold nanoparticles are used to stabilize the boundary of an inverse micelle system which contains an aqueous fluorescent dye solution on it's interior. A temperature dependent fluorescence intensity allows us to probe the temperature changes induced by laser irradiation. In the remainder of this thesis we describe a separate experiment involving the use of gold nanoparticles to study the free surface dynamics of thin glassy films. There is a growing body of evidence in the literature that thin polymer films in the glassy state exhibit heterogeneous dynamics; specifically that the first few nanometers from an air-polymer interface exhibit enhanced mobility relative to the interior of the film. The underlying mechanism responsible for this enhanced mobility remains elusive, however some believe it to be a direct consequence of the polymeric nature of these films. We describe in detail an experiment aimed at addressing this concern. We deposit gold nanoparticles onto the surface of a molecular (non-polymeric) glassy film and monitor their behaviour upon heating using atomic force microscopy. Our results clearly show the existence of enhanced surface mobility in the system studied and provide strong evidence that enhanced surface mobility should be expected in all glass forming systems.

A new class of polymer stabilized gadolinium trifluoride nanoparticles (NPs) have been developed as contrast agents for magnetic resonance imaging (MRI) and computed tomography (CT), with potential long term goals in targeted imaging and anti-cancer therapy. The NPs are comprised of a 90/10 mixture of GdF3/EuF3 and are coated with linear polyacrylic acid (PAA) chains consisting of 25 repeating units. The resulting aggregates are stable in serum and possess unprecedented mass relaxivities [i.e. ~100-200 s-1(mg/mL)-1]. Electron microscopy images reveal various NP morphologies which depend on the exact synthesis protocol. These include highly cross-linked oblong clusters with 30-70 nm cross sections, extensively cross-linked aggregates with 100-300 nm cross sections, and distinct polymer stabilized nanocrystals with 50 nm diameters. Their application as contrast agents in T1-weighted MRI studies, CT imaging at various X-ray energies, and preliminary rat brain perfusion studies was also tested. NP contrast enhancement was compared to Gd-DPTA (Magnevist®) and iopramide (Ultravist 300®) to demonstrate their high contrasting properties and potential as multimodal contrast agents.

This work begins with chemotaxis studies involving Salmonella typhimurium. Known chemical attractants (ribose, aspartic acid, etc.) and repellents (nickel chloride, sodium acetate, etc.) were tested to direct bacteria swimming patterns. It was found that high concentrations of both attractant and repellent, approximately 10% chemical in deionized (DI) water, yielded better separation results than lower concentrations, such as 1% and .1% chemical in DI water. Utilizing these attractants or repellents appropriately can allow live bacteria to be directed in a desired manner in a microfluidic device, while dead bacteria, which yield no response, can be separated into a waste reservoir. Another important aspect of bacteria separation is preconcentration, or the process of concentrating bacteria in a usable amount of liquid for further analysis in a microfluidic device. This study introduces a method of capturing Salmonella typhimurium through the use of magnetic nanoparticles (MNPs) without functionalizing them with antibody or amine coatings. Based on the work by Deng et al., MNPs were prepared in various ways to alter their diameter and surface characteristics to achieve optimal bacteria capture efficiency. A capture efficiency of approximately 94% has been achieved by altering chemical quantities in the MNP fabrication process. A macro-scale flow cell prototype was designed and characterized in order to ‘clean’ large volumes of buffer and separate the bacteria-MNP aggregates through the use of a magnetic field. Finally, intervention of bacteria is a significant topic in food safety applications. This study utilizes Fe (III) to inhibit bacteria growth. This chemical was used in the presence of Salmonella, E. coli, Staphylococcus, and Pseudomonas. Further experiments were conducted with raw chicken and lettuce contaminated with Salmonella typhimurium. Using as little as .005M Fe III in DI water, up to 5 orders of magnitude reduction in bacteria growth was seen on test plates as compared to control plates.

There are no clinically approved drug delivery strategies designed for localized and sustained release to the injured spinal cord, two features which are heavily exploited in pre-clinical demonstrations of efficacy. We have previously shown that injection of drug loaded hydrogels into the intrathecal space is safe, minimally invasive, and drug release localized to the site of injection for up to one day. In the present work we developed a platform for sustained release from 1 to 28 days based on a physical gel of methylcellulose with hyaluronan and poly(lactic-co-glycolic acid) (PLGA) nanoparticles added as gelation agents. These composite hydrogels met the design criteria of injectability, fast gelation, minimal swelling, and 28 day stability. Sustained release of 6 therapeutic molecules from the composite was achieved by encapsulation in the particles or dissolution in the hydrogel. Release of PLGA encapsulated drugs from the composite was linear for 28 days. Drugs dissolved in the hydrogel were released by Fickian diffusion. The HAMC hydrogel/PLGA nanoparticle composite was delivered to uninjured and spinal cord injured rats and the animals monitored for 14 and 28 days respectively. The composite was well tolerated in the intrathecal space with no impact on motor function as determined by the BBB scale and minimal inflammation in both studies. No increase in reactive astrocytes or cavity volume was found in clip compression spinal cord injured rats, indicating that the composite did not affect these aspects of the secondary injury cascade. We then turned to sustained release of anti-NogoA, a promising neuroregenerative molecule typically delivered for 2 - 4 weeks. Formulations of anti-NogoA or a model IgG were prepared and release was demonstrated over 28 days in vitro. Bioactivity was assessed using a novel ELISA which utilized anti-NogoA / NogoA binding to detect only active antibody, advantageous because anti-NogoA release can now be easily optimized prior to in vivo studies of efficacy. The key features of current work are the development of an intrathecal drug delivery platform, demonstration of safety in a rat model, and formulation for use with anti-NogoA.

There are no clinically approved drug delivery strategies designed for localized and sustained release to the injured spinal cord, two features which are heavily exploited in pre-clinical demonstrations of efficacy. We have previously shown that injection of drug loaded hydrogels into the intrathecal space is safe, minimally invasive, and drug release localized to the site of injection for up to one day. In the present work we developed a platform for sustained release from 1 to 28 days based on a physical gel of methylcellulose with hyaluronan and poly(lactic-co-glycolic acid) (PLGA) nanoparticles added as gelation agents. These composite hydrogels met the design criteria of injectability, fast gelation, minimal swelling, and 28 day stability. Sustained release of 6 therapeutic molecules from the composite was achieved by encapsulation in the particles or dissolution in the hydrogel. Release of PLGA encapsulated drugs from the composite was linear for 28 days. Drugs dissolved in the hydrogel were released by Fickian diffusion. The HAMC hydrogel/PLGA nanoparticle composite was delivered to uninjured and spinal cord injured rats and the animals monitored for 14 and 28 days respectively. The composite was well tolerated in the intrathecal space with no impact on motor function as determined by the BBB scale and minimal inflammation in both studies. No increase in reactive astrocytes or cavity volume was found in clip compression spinal cord injured rats, indicating that the composite did not affect these aspects of the secondary injury cascade. We then turned to sustained release of anti-NogoA, a promising neuroregenerative molecule typically delivered for 2 - 4 weeks. Formulations of anti-NogoA or a model IgG were prepared and release was demonstrated over 28 days in vitro. Bioactivity was assessed using a novel ELISA which utilized anti-NogoA / NogoA binding to detect only active antibody, advantageous because anti-NogoA release can now be easily optimized prior to in vivo studies of efficacy. The key features of current work are the development of an intrathecal drug delivery platform, demonstration of safety in a rat model, and formulation for use with anti-NogoA.

The interaction between metal nanoparticles and small molecules has been investigated by FTIR and UV-visible absorption spectroscopy. Electrospray deposition into an argon matrix was chosen as the initial method. An electrospray metal source was tested in development stage. Both the formation of a stable corona discharge as well as a stable Taylor cone were successfully completed. Problems arose when the entire system was tested. It was determined that the vacuum was insufficient for the length of the flight path. Focus then shifted to nanoparticles in more conventional environments. Sol-gel encapsulated nanoparticles were generated, in the form of both monoliths as well as thin film coatings on silicon wafers. The gels were exposed to 1atm of carbon monoxide in a gas cell. The method encountered problems due to spectral interference from the matrix. The next attempt consisted of solution stabilized nanoparticles. The solution was exposed to various amounts of both ammonium sulphate and diethylamine. There was again the problem of solvent interference, even when attempting to observe the system using Raman spectroscopy. Finally, surface stabilized nanoparticles were generated, using 3-mercaptopropyltrimethoxysilane to adhere the particles to glass slides. While the coating was successfully applied to the glass slides, as confirmed with Raman spectroscopy, it was not possible to get the nanoparticles to adhere. Future outlook for this project is briefly reviewed. / Thesis (Master, Chemistry) -- Queen's University, 2009-06-26 10:30:58.295

Nanotechnology as a science is emerging rapidly. As materials are synthesized and utilized at the nanometer size scale, concerns of potential health and safety effects are arising. In an effort to elucidate the physicochemical characteristics of nanoparticles influential in toxicological studies, surface properties of metal oxide and carbonaceous nanoparticles were measured. These properties include zeta potential, dissolution and surface-bound chemical components. Subsequently, the role of these properties in oxidative stress was examined in vitro. This work identifies the influence that pH has on the zeta potential of nanoparticles. The zeta potential has the ability to alter colloidal stability, as the largest nanoparticle agglomerate is seen at or near the isoelectric point for each of the particles tested. Furthermore, it was observed that metal oxide nanoparticles which exhibit a charged surface at physiological pH, lead to decreased in vitro cellular viability as compared to those that were neutral. Thus, nanoparticle zeta potential may be an important factor to consider when attempting to predict nanoparticle toxicity. Real world exposure to nanoparticles is a mixture of various particulates and organics. Therefore, to simulate this particle mixture, iron oxide (Fe2O3) and engineered carbon black (ECB) were utilized in combination to identify potential synergistic reactions. Following in vitro exposure, both nanoparticle types are internalized into endosomes, where liberated Fe3+ reacts with hydroquinone moieties on the ECB surface yielding Fe2+. This bioavailable iron may then generate oxidative stress through intracellular pathways including the Fenton reaction. As oxidative stress is common in particulate toxicology, a comparison between the antioxidant defenses of epithelial (A549) and mesothelial (MeT-5A) cell lines was made. The A549 cell line exhibits alterations in the NRF2-KEAP1 transcription factor system and therefore retains high basal levels of phase II antioxidants. Both cell types were exposed to 33 nm silica where intracellular oxidant generation coupled with markers of oxidative stress were observed. While the MeT-5A cells exhibited a decrease in cell viability, the A549 cell line did not. Therefore, proper characterization of both material and biological systems prior to toxicity testing will help to further define the risks associated with the use of nanotechnology.

Noble metal nanoparticles are of great interest due to their tunable optical and radiative properties. The specific wavelength of light at which the localized surface plasmon resonance occurs is dependent upon the shape, size and composition of the particle as well as the dielectric constant of the host medium. Thus, the optical properties of noble metal nanoparticles can be systematically tuned by altering these specific parameters. The purpose of this thesis is to investigate some of these properties related to metallic nanoparticles. The first several chapters focus on theoretical modeling to predict and explain various plasmonic properties of gold and silver nanoparticles while the later chapters focus on more accurately combining experimental and theoretical methods to explain the plasmonic properties of hollow gold nanoparticles of various shapes. The appendix contains a detailed description of the theoretical methods used throughout the thesis. It is intended to serve as a guide such that a user could carry out the various types of calculations discussed in this thesis simply by reading this appendix.